Cui Xiangyan, Fang Ning, Cui Yu, Xiao Dong, Wang Xin
Department of Otolaryngology-Head and Neck Surgery, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China.
Oncol Lett. 2019 Jun;17(6):4928-4934. doi: 10.3892/ol.2019.10150. Epub 2019 Mar 15.
The present study aimed to investigate the involvement of the recently identified long non-coding RNA neighboring enhancer of FOXA2 (lncRNA NEF) in laryngeal squamous cell carcinoma (LSCC). In this study, the expression levels of lncRNA NEF in tumor tissues and paired adjacent normal tissues were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Serum levels of NEF in patients with LSCC and healthy controls were also measured using RT-qPCR. Clinical and follow-up data of patients with LSCC were retrospectively analyzed. Diagnostic and prognostic values of serum NEF were evaluated by receiver operating characteristic curve and survival curve analysis, respectively. In addition, a NEF expression vector was constructed and transfected into human LSCC cells. The effects of NEF overexpression on cell proliferation, apoptosis and β-catenin expression were explored by Cell Counting kit-8 cell proliferation assay, MTT assay and western blotting. NEF was significantly downregulated in tumor tissues compared with in paired adjacent normal tissues of patients with LSCC. Serum levels of NEF were significantly lower in patients with LSCC than in healthy controls. Low serum levels of NEF distinguished patients with LSCC from healthy controls, and also indicated shorter postoperative survival. NEF overexpression inhibited proliferation and promoted apoptosis of LSCC cells, and also downregulated β-catenin expression. No significant effects of Wnt agonist on NEF expression were identified; however, Wnt agonist reduced the effects of NEF overexpression on cancer cell proliferation and apoptosis. In conclusion, lncRNA NEF may inhibit proliferation and promote apoptosis of LSCC cells by inhibiting Wnt/β-catenin signaling.
本研究旨在探讨最近鉴定出的叉头框蛋白A2（FOXA2）邻近增强子长链非编码RNA（lncRNA NEF）在喉鳞状细胞癌（LSCC）中的作用。在本研究中，通过逆转录定量聚合酶链反应（RT-qPCR）检测肿瘤组织和配对的相邻正常组织中lncRNA NEF的表达水平。还使用RT-qPCR测量LSCC患者和健康对照者血清中NEF的水平。对LSCC患者的临床和随访数据进行回顾性分析。分别通过受试者工作特征曲线和生存曲线分析评估血清NEF的诊断和预后价值。此外，构建NEF表达载体并将其转染到人LSCC细胞中。通过细胞计数试剂盒-8细胞增殖试验、MTT试验和蛋白质印迹法探讨NEF过表达对细胞增殖、凋亡和β-连环蛋白表达的影响。与LSCC患者配对的相邻正常组织相比，肿瘤组织中NEF显著下调。LSCC患者血清中NEF水平显著低于健康对照者。血清NEF水平低可将LSCC患者与健康对照者区分开来，也表明术后生存期较短。NEF过表达抑制LSCC细胞的增殖并促进其凋亡，还下调β-连环蛋白的表达。未发现Wnt激动剂对NEF表达有显著影响；然而，Wnt激动剂减弱了NEF过表达对癌细胞增殖和凋亡的影响。总之，lncRNA NEF可能通过抑制Wnt/β-连环蛋白信号通路来抑制LSCC细胞的增殖并促进其凋亡。
